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. 2001 May 15;98(11):6074–6079. doi: 10.1073/pnas.111114298

Figure 2.

Figure 2

Gab1 and the MBD of Gab1 form a complex with FRS2α, Shc, or Shp2 in cells expressing activated FGF receptor. 293 cells were transfected with expression vectors for kinase-inactive FGFR1 (KD) mutant, wt FGFR1, and FRS2α, Shc, or Shp2 (A–C) as indicated. Cell lysates were incubated with immobilized GST fusion proteins of full-length Gab1 (G) or the MBD of Gab1 (M). GST alone was used as a control (C). Bound proteins were eluted and analyzed by SDS–PAGE and immunoblotting with FRS2, Grb2, Shc, Shp2 antibodies (A–C). (D) Quiescent PC12 cells were unstimulated or stimulated with FGF1 and heparin (100 ng/ml and 5 μg/ml, respectively) or EGF (50 ng/ml) for 10 min. The lysates were immunoprecipitated with anti-Gab1 antibodies, resolved by SDS–PAGE, and followed by immunoblotting with anti-pTyr, Gab1, Grb2, FRS2, or Shp2 antibodies.